Arf6-dependent intracellular trafficking of pasteurella multocida toxin and pH-dependent translocation from late endosomes

Tana L. Repella, Mengfei Ho, Tracy P M Chong, Yuka Bannai, Brenda A. Wilson

Research output: Contribution to journalReview article

Abstract

The potent mitogenic toxin from Pasteurella multocida (PMT) is the major virulence factor associated with a number of epizootic and zoonotic diseases caused by infection with this respiratory pathogen. PMT is a glutamine-specific protein deamidase that acts on its intracellular G-protein targets to increase intracellular calcium, cytoskeletal, and mitogenic signaling. PMT enters cells through receptor-mediated endocytosis and then translocates into the cytosol through a pH-dependent process that is inhibited by NH4Cl or bafilomycin A1. However, the detailed mechanisms that govern cellular entry, trafficking, and translocation of PMT remain unclear. Co-localization studies described herein revealed that while PMT shares an initial entry pathway with transferrin (Tfn) and cholera toxin (CT), the trafficking pathways of Tfn, CT, and PMT subsequently diverge, as Tfn is trafficked to recycling endosomes, CT is trafficked retrograde to the ER, and PMT is trafficked to late endosomes. Our studies implicate the small regulatory GTPase Arf6 in the endocytic trafficking of PMT. Translocation of PMT from the endocytic vesicle occurs through a pH-dependent process that is also dependent on both microtubule and actin dynamics, as evidenced by inhibition of PMT activity in our SRE-based reporter assay, with nocodazole and cytochalasin D, respectively, suggesting that membrane translocation and cytotoxicity of PMT is dependent on its transfer to late endosomal compartments. In contrast, disruption of Golgi-ER trafficking with brefeldin A increased PMT activity, suggesting that inhibiting PMT trafficking to non-productive compartments that do not lead to translocation, while promoting formation of an acidic tubulovesicle system more conducive to translocation, enhances PMT translocation and activity.

Original languageEnglish (US)
Pages (from-to)218-241
Number of pages24
JournalToxins
Volume3
Issue number3
DOIs
StatePublished - Mar 1 2011

Fingerprint

Pasteurella multocida
Cholera Toxin
Endosomes
Transferrin
Nocodazole
Brefeldin A
Cytochalasin D
GTP Phosphohydrolases
Virulence Factors
Pathogens
Cytotoxicity
Glutamine
GTP-Binding Proteins
Recycling
Actins
Assays
Calcium
Membranes
Pasteurella multocida toxin
Proteins

Keywords

  • Endocytosis
  • Intoxication
  • Translocation

ASJC Scopus subject areas

  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Arf6-dependent intracellular trafficking of pasteurella multocida toxin and pH-dependent translocation from late endosomes. / Repella, Tana L.; Ho, Mengfei; Chong, Tracy P M; Bannai, Yuka; Wilson, Brenda A.

In: Toxins, Vol. 3, No. 3, 01.03.2011, p. 218-241.

Research output: Contribution to journalReview article

Repella, Tana L. ; Ho, Mengfei ; Chong, Tracy P M ; Bannai, Yuka ; Wilson, Brenda A. / Arf6-dependent intracellular trafficking of pasteurella multocida toxin and pH-dependent translocation from late endosomes. In: Toxins. 2011 ; Vol. 3, No. 3. pp. 218-241.
@article{e21e74777b5845e59aff9e60362540f5,
title = "Arf6-dependent intracellular trafficking of pasteurella multocida toxin and pH-dependent translocation from late endosomes",
abstract = "The potent mitogenic toxin from Pasteurella multocida (PMT) is the major virulence factor associated with a number of epizootic and zoonotic diseases caused by infection with this respiratory pathogen. PMT is a glutamine-specific protein deamidase that acts on its intracellular G-protein targets to increase intracellular calcium, cytoskeletal, and mitogenic signaling. PMT enters cells through receptor-mediated endocytosis and then translocates into the cytosol through a pH-dependent process that is inhibited by NH4Cl or bafilomycin A1. However, the detailed mechanisms that govern cellular entry, trafficking, and translocation of PMT remain unclear. Co-localization studies described herein revealed that while PMT shares an initial entry pathway with transferrin (Tfn) and cholera toxin (CT), the trafficking pathways of Tfn, CT, and PMT subsequently diverge, as Tfn is trafficked to recycling endosomes, CT is trafficked retrograde to the ER, and PMT is trafficked to late endosomes. Our studies implicate the small regulatory GTPase Arf6 in the endocytic trafficking of PMT. Translocation of PMT from the endocytic vesicle occurs through a pH-dependent process that is also dependent on both microtubule and actin dynamics, as evidenced by inhibition of PMT activity in our SRE-based reporter assay, with nocodazole and cytochalasin D, respectively, suggesting that membrane translocation and cytotoxicity of PMT is dependent on its transfer to late endosomal compartments. In contrast, disruption of Golgi-ER trafficking with brefeldin A increased PMT activity, suggesting that inhibiting PMT trafficking to non-productive compartments that do not lead to translocation, while promoting formation of an acidic tubulovesicle system more conducive to translocation, enhances PMT translocation and activity.",
keywords = "Endocytosis, Intoxication, Translocation",
author = "Repella, {Tana L.} and Mengfei Ho and Chong, {Tracy P M} and Yuka Bannai and Wilson, {Brenda A.}",
year = "2011",
month = "3",
day = "1",
doi = "10.3390/toxins3030218",
language = "English (US)",
volume = "3",
pages = "218--241",
journal = "Toxins",
issn = "2072-6651",
publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",
number = "3",

}

TY - JOUR

T1 - Arf6-dependent intracellular trafficking of pasteurella multocida toxin and pH-dependent translocation from late endosomes

AU - Repella, Tana L.

AU - Ho, Mengfei

AU - Chong, Tracy P M

AU - Bannai, Yuka

AU - Wilson, Brenda A.

PY - 2011/3/1

Y1 - 2011/3/1

N2 - The potent mitogenic toxin from Pasteurella multocida (PMT) is the major virulence factor associated with a number of epizootic and zoonotic diseases caused by infection with this respiratory pathogen. PMT is a glutamine-specific protein deamidase that acts on its intracellular G-protein targets to increase intracellular calcium, cytoskeletal, and mitogenic signaling. PMT enters cells through receptor-mediated endocytosis and then translocates into the cytosol through a pH-dependent process that is inhibited by NH4Cl or bafilomycin A1. However, the detailed mechanisms that govern cellular entry, trafficking, and translocation of PMT remain unclear. Co-localization studies described herein revealed that while PMT shares an initial entry pathway with transferrin (Tfn) and cholera toxin (CT), the trafficking pathways of Tfn, CT, and PMT subsequently diverge, as Tfn is trafficked to recycling endosomes, CT is trafficked retrograde to the ER, and PMT is trafficked to late endosomes. Our studies implicate the small regulatory GTPase Arf6 in the endocytic trafficking of PMT. Translocation of PMT from the endocytic vesicle occurs through a pH-dependent process that is also dependent on both microtubule and actin dynamics, as evidenced by inhibition of PMT activity in our SRE-based reporter assay, with nocodazole and cytochalasin D, respectively, suggesting that membrane translocation and cytotoxicity of PMT is dependent on its transfer to late endosomal compartments. In contrast, disruption of Golgi-ER trafficking with brefeldin A increased PMT activity, suggesting that inhibiting PMT trafficking to non-productive compartments that do not lead to translocation, while promoting formation of an acidic tubulovesicle system more conducive to translocation, enhances PMT translocation and activity.

AB - The potent mitogenic toxin from Pasteurella multocida (PMT) is the major virulence factor associated with a number of epizootic and zoonotic diseases caused by infection with this respiratory pathogen. PMT is a glutamine-specific protein deamidase that acts on its intracellular G-protein targets to increase intracellular calcium, cytoskeletal, and mitogenic signaling. PMT enters cells through receptor-mediated endocytosis and then translocates into the cytosol through a pH-dependent process that is inhibited by NH4Cl or bafilomycin A1. However, the detailed mechanisms that govern cellular entry, trafficking, and translocation of PMT remain unclear. Co-localization studies described herein revealed that while PMT shares an initial entry pathway with transferrin (Tfn) and cholera toxin (CT), the trafficking pathways of Tfn, CT, and PMT subsequently diverge, as Tfn is trafficked to recycling endosomes, CT is trafficked retrograde to the ER, and PMT is trafficked to late endosomes. Our studies implicate the small regulatory GTPase Arf6 in the endocytic trafficking of PMT. Translocation of PMT from the endocytic vesicle occurs through a pH-dependent process that is also dependent on both microtubule and actin dynamics, as evidenced by inhibition of PMT activity in our SRE-based reporter assay, with nocodazole and cytochalasin D, respectively, suggesting that membrane translocation and cytotoxicity of PMT is dependent on its transfer to late endosomal compartments. In contrast, disruption of Golgi-ER trafficking with brefeldin A increased PMT activity, suggesting that inhibiting PMT trafficking to non-productive compartments that do not lead to translocation, while promoting formation of an acidic tubulovesicle system more conducive to translocation, enhances PMT translocation and activity.

KW - Endocytosis

KW - Intoxication

KW - Translocation

UR - http://www.scopus.com/inward/record.url?scp=79953245323&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79953245323&partnerID=8YFLogxK

U2 - 10.3390/toxins3030218

DO - 10.3390/toxins3030218

M3 - Review article

C2 - 22053287

AN - SCOPUS:79953245323

VL - 3

SP - 218

EP - 241

JO - Toxins

JF - Toxins

SN - 2072-6651

IS - 3

ER -